ES2566787T3 - Liquid pharmaceutical composition comprising nitisinone - Google Patents

Abstract

A liquid pharmaceutical formulation suitable for oral administration, comprising (a) a suspension of an effective amount of micronized 2- (2-nitro-4-trifluoromethylbenzoyl) -1,3-cyclohexanedione (nitisinone); and (b) citric acid buffer having a pH in the range of 2.5 to 3.5, preferably pH 3.0.

Description

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DESCRIPTION

Ifquida pharmaceutical composition comprising nitisinone TECHNICAL FIELD

This invention relates to pharmaceutical formulations comprising 2- (2-nitro-4-trifluoromethylbenzoyl) -1,3-cyclohexanedione (nitisinone) as an active agent. The formulations are useful in the treatment of disorders and diseases where inhibition of 4-hydroxyphenylpyruvate dioxygenase (HPPD) is desirable, for example, in hereditary type I tyrosinemia.

PREVIOUS TECHNIQUE

The compound 2- (2-nitro-4-trifluoromethylbenzoyl) -1,3-cyclohexanedione, also known as nitisinone or NTBC, was first described as a herbicide (US 5,006,158; 4,695,673; 5,668. 089).

Nitisinone is used under the trade name Orfadin® for the treatment of hereditary tyrosinemia type I (HT-1), a rare pediatric disease. HT-1 is a genetic metabolic disorder that is the result of an inability to break down the amino acid tyrosine. Because it causes liver failure and liver cancer, children with HT-1 rarely live more than twenty years without a liver transplant.

As described, for example, in US 5,550,165, nitisinone is a competitive inhibitor of 4-hydroxyphenyl-pyruvate dioxygenase (HPPD), an enzyme prior to smokingylacetoacetate (FAH) hydrolase ) on the catabolic route of tyrosine. By inhibiting the normal catabolism of tyrosine in patients with HT-1, nitisinone prevents the accumulation of malebolic acetoacetate and fumaryl acetoacetate catabolic intermediates. In patients with HT-1, these catabolic intermediates are converted into the toxic metabolites succinylacetone and succinylacetoacetate, which are responsible for the toxicity observed in the liver and kidney.

In addition, nitisinone has been described as useful in the treatment of other disorders, such as Parkinson's disease (WO 2006/090117); depression (WO 2008/020150); the smdrome of the restless leg (document WO 2010/054273); and alcaptonuria (Sunwanarat, P. et al., Metabolism 54: 719-728, 2005). The use of nitisinone has also been described in a method to improve phagolysosomal fusion after infection of a patient with a microorganism (US patent application document, publication No. 20100227936).

Oral administration of drugs is one of the preferred routes for treatment, due to its simplicity. Although the drugs are generally administered in the form of tablets or capsules, such administration may be less preferred, for example, when the dosage has to be accurately adapted to the treated subject, or it may be less convenient, for example, in the case of Pediatric or veterinary drugs. The liquid dosage form may then be an advantageous alternative.

Therefore, there is a need for stable liquid nitisinone compositions that are adapted for administration to pediatric patients and to overcome the drawbacks with solid pharmaceutical compositions.

DESCRIPTION OF THE INVENTION

According to the invention, it has been shown that a liquid pharmaceutical formulation, comprising a suspension of micronized nitisinone and having a pH of about 3, has surprisingly advantageous properties, such as increased stability. Accordingly, the present invention relates to a liquid pharmaceutical formulation suitable for oral administration, comprising (a) a suspension of an effective amount of 2- (2-nitro-4-trifluoromethylbenzoyl) -1,3-cyclohexanedione (nitisinone) micronized; and (b) a citric acid buffer having a pH in the range of 2.5 to 3.5, preferably pH 3.0.

The expression "effective amount" of nitisinone should be understood as an amount effective to inhibit 4-hydroxyphenylpyruvate dioxygenase. Preferably, the amount of nitisinone is 1 to 10 mg / ml, more preferably 4 mg / ml.

Nitisinone can be obtained by conventional organic chemistry procedures that are already known for the production of materials of analogous structure. Thus, for example, nitisinone can be conveniently obtained by reacting 2-nitro-4-trifluoromethylbenzoyl chloride with cyclohexane-1,3-dione in the presence of acetone cyanohydrin and a suitable base, such as triethylamine, as described in the US document 5,550,165. The starting 2-nitro-4-trifluoromethylbenzoyl chloride can be obtained from the corresponding benzoic acid, for example, by a reaction with thionyl chloride or oxalyl chloride, as described in Reagents for Organic Synthesis, (J Wiley and Sons, 1967; vol. 1, pages 767-769) and is generally used without special purification. Similarly, 2-nitro-4-trifluroromethylbenzoic acid can be obtained, for example, as described in Haupstein et al. in J. Amer. Chem. Soc., 1954, 76, 1051, or by one of the general methods described in The Chemistry of Carboxylic Acids and Esters (J Wiley and Sons, 1969; compiler: S. Patai) and Survey of Organic Synthesis (J Wiley and Sons, 1970; CA Buehler and DF Pearson).

Preferably, the formulation according to the invention further comprises one or more pharmaceutically acceptable components, selected from the group consisting of suspending agents, sweeteners, preservatives, surfactants and flavoring agents.

A suitable suspending agent is, for example, hydroxypropyl methylcellulose (HPMC) in an amount of 1 to 20 mg / ml, preferably 5 mg / ml.

A suitable sweetener is glycerol, in an amount that results in an acceptable taste. The amount of glycerol is preferably 100 to 500 mg / ml, more preferably 500 mg / ml.

The formulation according to the invention preferably comprises at least one preservative selected from methyl paraben, propyl paraben and sodium benzoate. Preferably, the preservatives are methyl paraben 10 in an amount of 1 to 2 mg / ml, more preferably 1.4 mg / ml; propyl paraben in an amount of 0.1 to 0.2 mg / ml, more preferably 0.14 mg / ml; and sodium benzoate in an amount of 0.2 to 5 mg / ml, more preferably 1.0 mg / ml.

The formulation according to the invention preferably comprises a surfactant, such as polysorbate 80 (polyoxyethylene sorbitan monooleate (80); common trademarks include Alkest TW 80® and 15 Tween 80®). The amount of polysorbate 80 should be sufficient to moisten the nitisinone particles to facilitate the dispersion of the nitisinone during manufacturing, as well as to avoid any agglomeration of the nitisinone particles during storage of the final product. Preferably, the formulation according to the invention comprises polysorbate 80 in an amount of 0.1 to 20 mg / ml, more preferably 0.10 to 0.15 mg / ml, such as about 0.135 mg / ml.

The formulation according to the invention preferably comprises an aromatic agent, such as strawberry flavor. The amount of flavoring agent should be sufficient to achieve an acceptable taste of the formulation and preferably in an amount of 0.2 to 1.1 mg / ml, more preferably 0.7 mg / ml.

In a particularly preferred form, the formulation according to the invention comprises

(a) nitisinone (4 mg / ml);

25 (b) cfric acid monohydrate (9 mg / ml);

(c) trisodium citrate dihydrate (2.1 mg / ml)

(d) hydroxypropyl methylcellulose (5 mg / ml);

(e) glycerol (500 mg / ml);

(f) methyl paraben (1.4 mg / ml);

30 (g) propyl paraben (0.14 mg / ml); Y

(h) polysorbate 80 (0.14 mg / ml).

In another especially preferred form, the formulation according to the invention comprises

(a) nitisinone (4 mg / ml);

(b) cfric acid monohydrate (9 mg / ml);

35 (c) trisodium citrate dihydrate (2.1 mg / ml)

(d) hydroxypropyl methylcellulose (5 mg / ml);

(e) glycerol (500 mg / ml);

(f) sodium benzoate (1.0 mg / ml); Y

(g) polysorbate 80 (0.14 mg / ml).

An additional preferred form of the formulation comprises a flavoring agent such as:

(h) strawberry flavor (0.7 mg / ml).

The formulation according to the invention is useful for the treatment of medical disorders and diseases in which the inhibition of 4-hydroxyphenyl pyruvate dioxygenase (HPPD) is desirable. Examples of such conditions include hereditary tyrosinemia type 1 (HT-1), Parkinson's disease, depression, restless leg smdrome and alkaptonuria.

The formulation according to the invention is particularly useful for pediatric use. Specifically, it is suitable for newborns and children up to 8-10 years old, representing a body weight range of approximately 3.5 to 40 kg. A daily dose of 1 mg / kg therefore corresponds to a dose range of 2 x 1.75 mg to 2 x 20 mg. A presentation of 4 mg / ml will achieve acceptable dosage volumes, corresponding 0.44 5 to 5 ml administered twice a day. An oral syringe is suitable as an administration dispenser for precise dosing in this interval.

EXAMPLES

EXAMPLE 1: Micronization of nitisinone

A laboratory-scale air jet mill, a 2-inch evaluation model of Sturtevant Inc., 10 was used to micronize the nitisinone, obtained from Bachem, Switzerland. The mill was operated with tangential flow (ie, the air and the drug are fed in the same direction in the grinding chamber). The unmilled drug was introduced into the mill using a Venturi feed system, Syncron®, F-TO-C magnet magnet model, in which air is used to attract the feed material to the grinding chamber. A product filter bag was fixed at the outlet of the mill, through which the exhaust gases and the ground drug 15 were collected. The milling conditions were established as follows:

• Grinding air: dry nitrogen gas

• Grinding pressure: 6.20 bar (90 psi)

• Feed pressure: 5.86 bar (85 psi)

• Working conditions: Environmental

20 5 g of the API was passed through the laboratory scale micronizer and the resulting material was collected (3.7 g). The material was analyzed for / by the particle size diameter (PSD) diameter test, and high performance liquid chromatography purity (HPLC), powder X-ray diffraction (XRPD), differential scanning calorimetry (DSC ) and polarized light microscopy (PLM). The results of the micronization pSd are shown in Table I.

Table I

Diameter of particle size (microns)

 d-ici d20 d50 d80 d90

 Starting material

 20.50 33.10 60.01 94.42 115.11

 Micronized material

 0.30 0.47 1.29 2.59 3.59

25

EXAMPLE 2: Preparation of an oral suspension of micronized nitisinone containing methyl paraben and propyl paraben as preservatives

A formulation according to the invention, as shown in Table II, was prepared according to conventional procedures.

30 Table II

 Ingredient

 Amount (mg) Function

 Nitisinone (micronized)

 4.0 Active substance

 Hydroxypropyl methylcellulose (HPMC)

 5.0 Suspension Agent

 Glycerol

 500 Sweetener

 Polysorbate 80

 0.135 Surfactant

 Methyl Paraben

 1.4 Preservatives

 Propyl paraben

 0.14

 Dric acid monohydrate

 8.98 Buffer (pH 3.0)

 Trisodium citrate dihydrate

 2.13

 Purified water

 c.s.p. up to 1.00 ml Solvent

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EXAMPLE 3: Preparation of an oral suspension of micronized nitisinone containing sodium benzoate as a preservative and strawberry aroma as a flavoring agent

A formulation according to the invention, as shown in Table III, was prepared according to conventional procedures.

Table III

 Ingredient

 Amount (mg) Function

 Nitisinone (micronized)

 4.0 Active substance

 Hydroxypropyl methylcellulose (HPMC)

 5.0 Suspension Agent

 Glycerol

 500 Sweetener

 Polysorbate 80

 0.135 Surfactant

 Sodium benzoate

 1.0 Preservative

 Strawberry aroma

 0.7 Savoring agent

 Cfric acid monohydrate

 8.98 Buffer (pH 3.0)

 Trisodium citrate dihydrate

 2.13

 Purified water

 c.s.p. up to 1.00 ml Solvent

EXAMPLE 4: Preparation of a solution of nitisinone for comparison

A solution of nitisinone, as shown in Table IV, was prepared according to conventional procedures.

Table IV

 Ingredient

 Amount (mg) Function

 Nitisinone

 2.0 Active substance

 Methyl Paraben

 1.8 preservatives

 Propyl paraben

 0.2

 KH2PO4

 1.4 Buffer (pH 6.8)

 Na2HPO42H2O

 2.9

 NaOH (0.5 M aqueous)

 Adjust to pH 6.8

 Purified water

 c.s.p. up to 1.00 ml Solvent

EXAMPLE 5: Optimization of preservative quantities through microbial exposure studies according to the European Pharmacopoeia (Ph Eur 5.1.3) and the United States Pharmacopoeia (uSp <51>)

The results of different amounts of preservative are shown in Tables V and VI, below.

Table V

 Oral suspension of micronized nitisinone prepared according to Example 2 containing different amounts of methyl paraben and propyl paraben as preservatives.

 Methyl paraben / Propyl paraben (mg / mL) Limits

 Microbe

 Days 0 1.0 / 0.1 1.4 / 0.14 1.7 / 0.17 2.0 / 0.2 Ph Eur 5.1.3 USP <51> Units

 S. aureus

 Initial 5.3-5.5 5.3-5.5 5.3-5.5 5.3-5.5 5.3-5.5 - - log

 14

 > 3.5> 3.5> 3.5> 3.3> 3.5> 3> 1.0 log network

 28

 YES YES YES YES YES YES log red

 P. aeruginosa

 Initial 5.3-5.5 5.3-5.5 5.3-5.5 5.3-5.5 5.3-5.5 - - log

 Oral suspension of micronized nitisinone prepared according to Example 2 containing different amounts of methyl paraben and propyl paraben as preservatives.

 Methyl paraben / Propyl paraben (mg / mL) Limits

 Microbe

 Days 0 1.0 / 0.1 1.4 / 0.14 1.7 / 0.17 2.0 / 0.2 Ph Eur 5.1.3 USP <51> Units

 14

 > 3.3> 3.5> 3.5> 3.4> 3.5> 3> 1.0 log network

 28

 YES YES YES YES YES YES log red

 E. coli

 Initial 5.2-5.6 5.2-5.6 5.2-5.6 5.2-5.6 5.2-5.6 - - log

 14

 > 3.7> 3.6> 3.6> 3.2> 3.6> 3> 1.0 log network

 28

 YES YES YES YES YES YES log red

 C. albicans

 Initial 5.3-5.6 5.3-5.6 5.3-5.6 5.3-5.6 5.3-5.6 - - log

 14

 1.4> 3.7> 3.7> 3.3> 3.7> 1 SI log network

 28

 2.2 YES YES YES YES YES YES log network

 A. brasiliensis

 Initial 5.5-5.6 5.5-5.6 5.5-5.6 5.5-5.6 5.5-5.6 - - log

 14

 1.0 2.2 2.1 3.2> 3.6> 1 SI log network

 28

 1.0 YES 3.3 3.3 YES YES YES log network

 YES = No increase

Table VI

Oral suspension of micronized nitisinone prepared according to Example 3 containing different amounts of sodium benzoate as a preservative.

 Sodium Benzoate (mg / mL) Limits

 Microbe

 Days 0.2 1.0 3.0 5.0 Ph Eur 5.1.3 USP <51> Units

 S.aureus

 Initial 5.3 5.3 5.3 5.3 - - log

 14

 5 5 5 5> 3> 1 log network

 28

 YES YES YES YES YES YES log red

 P. aeruginosa

 Initial 5.2 5.2 5.2 5.2 - - log

 14

 5 5 5 5> 3> 1 log network

 28

 YES YES YES YES YES YES log red

 E. coli

 Initial 5.4 5.4 5.4 5.4 - - log

 14

 5 5 5 5> 3> 1 log network

 28

 YES YES YES YES YES YES log red

 C. albicans

 Initial 5.8 5.8 5.8 5.8 - - log

 14

 1.4 4.5 5 5> 1 YES log network

 28

 4.1 5 YES YES YES YES log network

 A. brasiliensis

 Initial 5.6 5.6 5.6 5.6 - - log

 14

 1 3,3 5 5> 1 YES log network

 28

 1.3 5 YES YES YES YES log network

YES = No increase

The results show that all the above formulations according to the invention meet the requirements set forth for preservative efficacy, in accordance with the European Pharmacopoeia (Ph Eur) and the United States Pharmacopoeia (USP), including formulation without preservatives, which indicates a self-serving nature

Vadora of the basic formulation.

EXAMPLE 6: Stability Test

Samples from the oral suspension of micronized nitisinone prepared according to Example 2, as well as the nitisinone solution prepared according to Example 4, were put in stability at + 5 ° C, + 25 ° C and + 40 ° C, res - 5 pectively, for l2 months. At the concentrations of nitisinone and product degradation product 6- (trifluoromethyl) -3,4-dihydro-1H-xanthene-1,9 (2H) -dione (oxotetrahydroxantenone) followed by HPLC with UV detection. The results, shown in Tables VII to X, are then expressed as a percentage of the nominal concentration of nitisinone (% of the label indication).

Table VII

 Oral suspension of micronized nitisinone prepared according to Example 2.

 Nitisinone (% of label indication)

 Temperature

 Months

 0

 1 2 3 6 12

 5 ° C

 99.9 104.2 101.7 105.0 102.9 104.8

 25 ° C

 99.9 105.6 98.6 104.0 101.8 103.7

 40 ° C

 99.9 105.6 102.0 102.7 101.0 100.1

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Table VIII

Oral suspension of micronized nitisinone prepared according to Example 2 (nd = not detected). Oxotetrahydroxantenone (% of label indication)

Months

 Temperature

 0 1 2 3 6 12

 5 ° C

 nd nd nd nd nd nd

 25 ° C

 nd nd nd nd 0.02 0.02

 40 ° C

 nd nd 0.07 0.15 0.28 0.54

Table IX

Nitisinone solution prepared according to Example 4. Nitisinone (% of label indication)

Months

 Temperature

 0 1 2 3 6 12

 5 ° C

 96.6 99.8 95.3 100.4 99.7 99.3

 25 ° C

 96.6 100.9 96.0 100.2 98.0 95.9

 40 ° C

 96.6 98.3 96.3 93.6 86.5 74.4

15 Table X

 Nitisinone solution prepared according to Example 4 (nd = not detected).

 Oxotetrahydroxantenone (% of label indication)

 Months

 Temperature

 0 1 2 3 6 12

 5 ° C

 nd nd nd nd 0.03 0.05

Nitisinone solution prepared according to Example 4 (nd = not detected). Oxotetrahydroxantenone (% of label indication)

Months

 Temperature

 0 1 2 3 6 12

 25 ° C

 nd 0.01 nd 0.39 0.58 0.78

 40 ° C

 nd 0.07 1.86 2.05 1.63 1.38

The results show that the formulation according to the invention (Tables VII and VIII) is more stable than the comparison solution (Tables IX and X) in all storage conditions. In the comparison solution, the main degradation product, oxotetrahydroxantenone, is further degraded to secondary degradation products. As a consequence, it is not possible to achieve a mass balance between nitisinone and degradation products for the reference solution.

EXAMPLE 7: Stability of oxotetrahydroxantenone

The study of the stability of the main degradation product in Example 6, oxotetrahydroxantenone, is carried out under conditions similar to those described in Example 6. The samples of oxotetrahydroxantenone (OTHX), 81 10 pg / ml, either in buffer citrate pH 3.0 or phosphate buffer pH 6.8, were put in stability at + 5 ° C, + 25 ° C and + 37 ° C, respectively, for 6 months. The concentrations of OTHX and the secondary degradation products 1,3-cyclohexanedione (CHD) and 4- (trifluoromethyl) salidic acid (TSA) were analyzed by LC-MS. The results, shown in Table XI, below, are expressed as the percentage of the initial OTHX concentration. The mass balance expressed as the total recovery of CHD + OTHX + TSA compared to the initial concentration of 15 OTHX, was calculated from MmOTHX / (MmCHD + MmTSA) x (CHDconc + TSAconc) + OTHXconc expressed in pg / ml , where MmOTHX, MmCHD and MmTSA are the molecular masses corresponding to 282, 202 and 206 g / mol, respectively. The results for mass balance, expressed as a percentage of the initial OTHX concentration, are shown in Table XII.

Table XI

Stability of oxotetrahydroxantenone solutions prepared according to Example 7.

 Citrate buffer pH 3.0 Phosphate buffer pH 6.8

 Months

 Months

 Component

 Temperature (° C) 1.8 3 6 1.8 3 6

 OTHX

 5 96.7 96.9 91.6 96.7 91.0 83.1

 25

 96.7 102.7 92.3 74.8 63.2 44.8

 37

 97.9 97.7 95.3 33.5 13.7 0.4

 CHD

 5 0.0 0.0 0.0 0.0 0.7 0.0

 25

 0.0 0.0 0.0 7.7 9.7 25.2

 37

 0.0 0.0 0.0 22.2 21.5 32.8

 TSA

 5 0.0 0.0 0.0 1.4 1.9 0.0

 25

 0.0 0.0 0.0 13.0 20.2 36.4

 37

 0.2 0.0 0.0 38.6 49.1 66.4

Table XII

 Balancing mass

 Citrate buffer pH 3.0 Phosphate buffer pH 6.8

 Months

 Months

 Temperature (° C)

 1.8 3 6 1.8 3 6

 5

 96.7 96.9 91.6 97.9 93.3 83.1

 Balancing mass

 Citrate buffer pH 3.0 Phosphate buffer pH 6.8

 Months

 Months

 25

 96.7 102.7 92.3 93.2 89.8 99.4

 37

 98.1 97.7 95.3 87.4 76.3 88.5

The results show that the formulation according to the invention is surprisingly stable also with respect to the formation of secondary degradation products. The results, close to 100% for mass equilibrium, confirm that the LC-MS method is capable of detecting and determining the mayone of the secondary degradation products.

Claims (18)

5 10 fifteen twenty 25 30 35 40 1. A pharmaceutical formula Kquida suitable for oral administration, comprising (a) a suspension of an effective amount of micronized 2- (2-nitro-4-trifluoromethylbenzoyl) -1,3-cyclohexanedione (nitisinone); Y (b) Cfric acid buffer having a pH in the range of 2.5 to 3.5, preferably pH 3.0. 2. The formulation according to claim 1, wherein the amount of nitisinone is 1 to 10 mg / ml, preferably 4 mg / ml. 3. The formulation according to claim 1, further comprising one or more pharmaceutically acceptable constituents selected from the group consisting of suspending agents, sweeteners, preservatives, surfactants and flavoring agents. 4. The formulation according to claim 3, wherein the suspending agent is hydroxypropyl methylcellulose. 5. The formulation according to claim 4, wherein the suspending agent is hydroxypropyl methylcellulose in an amount of 1 to 20 mg / ml, preferably 5 mg / ml. 6. The formulation according to claim 3, wherein the sweetener is glycerol. 7. The formulation according to claim 6, wherein the sweetener is glycerol in an amount of 100 to 500 mg / ml, preferably 500 mg / ml. 8. The formulation according to claim 3, wherein the preservative is methyl paraben and / or propyl paraben. 9. The formulation according to claim 8, wherein the preservatives are methyl paraben in an amount of 1 to 2 mg / ml, preferably 1.4 mg / ml, and propyl paraben in an amount of 0.1 to 0.2 mg / ml, preferably 0.14 mg / ml. 10. The formulation according to claim 3, wherein the preservative is sodium benzoate in an amount of 0.2 to 5 mg / ml, preferably 1 mg / ml. 11. The formulation according to claim 3, wherein the surfactant is polysorbate 80. 12. The formulation according to claim 11, wherein the surfactant is polysorbate 80 in an amount of 0.1 at 20 mg / ml, preferably 0.10 to 0.15 mg / ml. 13. The formulation according to any one of claims 1-9, 11 or 12, comprising: (a) nitisinone (4 mg / ml); (b) dric acid monohydrate (9 mg / ml); (c) trisodium citrate dihydrate (2.1 mg / ml) (d) hydroxypropyl methylcellulose (5 mg / ml); (e) glycerol (500 mg / ml); (f) methyl paraben (1.4 mg / ml); (g) propyl paraben (0.14 mg / ml); Y (h) polysorbate 80 (0.14 mg / ml). 14. The formulation according to any one of claims 1-7 or 10-12, comprising: (a) nitisinone (4 mg / ml); (b) cfric acid monohydrate (9 mg / ml); (c) trisodium citrate dihydrate (2.1 mg / ml) (d) hydroxypropyl methylcellulose (5 mg / ml); (e) glycerol (500 mg / ml); (f) sodium benzoate (1.0 mg / ml); Y (g) polysorbate 80 (0.14 mg / ml). 15. The formulation according to claim 13 or 14, further comprising a flavoring agent. 16. The formulation according to any one of claims 1 to 15, for use in the treatment of a medical condition selected from tyrosinemia, Parkinson's disease, depression, restless leg syndrome 5 ta and alcaptonuria. 17. The formulation according to claim 16, for use in the treatment of hereditary tyrosinemia type 1 (HT-1). 18. The formulation according to claim 17, for use in the treatment of hereditary tyrosinemia type 1 (HT-1) in a pediatric patient.